Anti-Nucleophosmin antibody [EP1848Y]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Nucleophosmin antibody [EP1848Y] (AB52644)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Nucleophosmin with purified ab52644 at 1/100 dilution (5 © : g/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor� 594) 1/200 (2.5 © : g/ml). Goat anti rabbit IgG (Alexa Fluor� 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 © : g/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nucleophosmin antibody [EP1848Y] (AB52644)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling Nucleophosmin with purified ab52644 at 1/300 dilution (1.14 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Nucleophosmin antibody [EP1848Y] (AB52644)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells, labeling Nucleophosmin with Purified ab52644 at 1/40 dilution (10μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Lab

Western blot - Anti-Nucleophosmin antibody [EP1848Y] (AB52644)

All lanes:

Western blot - Anti-Nucleophosmin antibody [EP1848Y] (ab52644) at 1/500000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MEF (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

3T3-L1 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 37 kDa

false

• WB

Unknown

Western blot - Anti-Nucleophosmin antibody [EP1848Y] (AB52644)

The band at 250kDa might be an oligomer and the band at 33kDa might be an isoform of NPM.

Blocking/Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Nucleophosmin antibody [EP1848Y] (ab52644) at 1/2000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 250 kDa,27 kDa,33 kDa

false